Description Lambda is a temperate Escherichia coli bacteriophage. The virion DNA is linear and double-stranded (48502 nt) with 12 bp single-stranded complementary 5’-ends. After the phage particle injects its chromosome into the cell, the chromosome circularizes by end joining. In the lytic pathway, phage genes encoding replication, lysis and virion proteins are expressed. The chromosome replicates and the replicas are cleaved and packaged into progeny phage particles. In the lysogenic pathway, phage gene expression is repressed, and the circular chromosome inserts into the bacterial chromosome by recombination. Phage lambda DNA is a common substrate for restriction endonucleases and for gnerating DNA size marker fragments. For large scale isolation of phage DNA, cI857Sam7, a mutant carrying four known mutations, is often used. The DNA sequence used to construct a phage alpha restriction map includes these mutations. The map shows enzymes that cut lambda DNA once. Enzymes produced by Fermentas are shown in orange. GenBank/EMBL Accession Numbers J02459, M17233, M24325, V00636, X00906. Applications Activity and specificity assays of restriction enzymes sensitive to Dam or Dcm methylation. Preparation of DNA molecular weight standards. Cloning. Quality Control DNA concentration is confirmed spectrophotometrically. Identity and homogenity of DNA is confirmed by digestion by BclI and CfrI, and analysis of DNA fragmentation patterns. |